Stretched exponential decay and correlations in the catalytic activity of fluctuating single lipase molecules

Ophir Flomenbom, Kelly Velonia, Davey Loos, Sadahiro Masuo, Mircea Cotlet, Yves Engelborghs, Johan Hofkens, Alan E. Rowan, Roeland J.M. Nolte, Mark Der Van Auweraer, Frans C. De Schryver, Joseph Klafter

Research output: Contribution to journalArticlepeer-review

Abstract

Single-molecule techniques offer a unique tool for studying the dynamical behavior of individual molecules and provide the possibility to construct distributions from individual events rather than from a signal stemming from an ensemble of molecules. In biological systems, known for their complexity, these techniques make it possible to gain insights into the detailed spectrum of molecular conformational changes and activities. Here, we report on the direct observation of a single lipase-catalyzed reaction for extended periods of time (hours), by using confocal fluorescence microscopy. When adding a profluorescent substrate, the monitored enzymatic activity appears as a trajectory of "on-state" and "off-state" events. The waiting time probability density function of the off state and the state-correlation function fit stretched exponentials, independent of the substrate concentration in a certain range. The data analysis unravels oscillations in the logarithmic derivative of the off-state waiting time probability density function and correlations between off-state events. These findings imply that the fluctuating enzyme model, which involves a spectrum of enzymatic conformations that interconvert on the time scale of the catalytic activity, best describes the observed enzymatic activity. Based on this model, values for the coupling and reaction rates are extracted.

Original languageEnglish
Pages (from-to)2368-2372
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume102
Issue number7
DOIs
StatePublished - 15 Feb 2005

Keywords

  • Single enzyme activity
  • Two-state trajectories

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